ArciTect™ CRISPR-Cas9 Genome Editing System

The ArciTect™ product family is a CRISPR-Cas9 genome editing system that uses ribonucleoprotein (RNP) complexes composed of purified Cas9 protein and custom synthetic guide RNA. Compared to previous technologies that utilize plasmid or mRNA-based systems, an RNP-based system enables efficient delivery and expression of CRISPR machinery in difficult-to-manipulate cells, including stem and primary cells. Once inside the cell, the RNP complexes will not induce the cellular immune response and will degrade in a timely manner to reduce off-target effects (Table 1). With validated reagents and protocols, the ArciTect™ system enables you to perform high-efficiency genome editing and generate functional gene-edited cells in your own lab.

Overcome Hurdles in Efficiency

Overcome challenges in efficient delivery and expression of CRISPR machinery using an RNP-based CRISPR-Cas9 system. ArciTect™ enables researchers to perform high-efficiency genome editing in difficult-to-manipulate cell types, including stem and primary cells.

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Why Use ArciTect™?

  • Maximize delivery and expression in difficult-to-manipulate cell types by using RNP complexes.
  • Simplify genome editing with an integrated guide RNA design tool and cell-type-specific protocols.
  • Get to your results faster with ready-to-use purified Cas9 proteins and synthetic guide RNAs.
  • Minimize potential off-target cutting with timely degradation of the RNP complex.

Figure 1. Genome Editing Workflow

Table 1. Comparison Between Different CRISPR Methods. 1


Genome Editing Protocols and Data

Explore step-by-step instructions for performing high-efficiency genome editing using CRISPR-Cas9 in a variety of cell lines, including stem and primary cell types. The protocols are optimized and validated with a case study and supporting data, and include important cell culture considerations and methods to evaluate editing efficiency.

Human Pluripotent Stem Cells (hPSCs)

Gene knockout and knock-in in hPSCs using electroporation and chemical transfection.
Case study: Knockout and Knock-in optimization through GFP to BFP conversion

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Human Primary T Cells

Gene knockout in primary human T cells using electroporation.
Case study: Evaluation of optimal culture methods for high-efficiency TRAC knockout

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CD34+ Human Hematopoietic Stem and Progenitor Cells (HSPCs)

Gene knockout in HSPCs using electroporation.
Case study: Evaluation of optimal culture methods for high-efficiency genome editing in CD34+ HSPCs

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Immunofluorescence image of a human adult stem cell-derived intestinal organoid.

Human Intestinal Organoids

Gene knockout in human adult stem cell-derived intestinal organoids using electroporation.

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Genome Editing Products

Guide RNA (gRNA)

Features:

  • Single guide RNA (sgRNA) containing the crRNA and tracrRNA regions within a single molecule
  • Two-part gRNA system composed of ArciTect™ crRNA and ArciTect™ tracrRNA
  • Compatible with all Cas9 nucleases

Applications:

  • Use ArciTect™ sgRNA or ArciTect™ crRNA and tracrRNA to guide the Cas9 nuclease to a specific target location

Cas9

Features:

  • RNP complex is active immediately following transfection
  • All versions of Cas9 contain nuclear localization signals for rapid translocation into the nucleus

Applications:

  • Use ArciTect™ Cas9 Nuclease to generate double-strand breaks at specific locations in the genome
  • Use ArciTect™ Cas9-eGFP Nuclease to optimize transfection conditions or sort cells following transfection

Positive Control Kits

Features:

  • ArciTect™ Human CRISPR Optimization Kits are flow cytometry-based kits designed to enable rapid optimization of genome editing in human cells
  • Validated for use with ArciTect™ family of products for genome editing

Applications:

  • ArciTect™ Human CRISPR Optimization Kit can be used to rapidly quantify genome editing efficiency, and can be used as a positive control
  • ArciTect™ Human HPRT Primer Mix can be used in a T7 endonuclease assay to assess cleavage efficiency
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CRISPR Design Tools

The CRISPR Guide Design Tool uses best practices and the latest computational tools to deliver the optimal CRISPR RNA (crRNA) or single guide RNA (sgRNA) sequence for every gene in the human and mouse genomes.


Scientific Resources

CRISPR-Cas9 Genome editing

Wiley E-book: Genome Editing Applications

Learn about next-generation disease modeling using CRISPR, including comprehensive genome editing strategies for complex cell culture models, suggestions for optimizing experimental conditions, and troubleshooting tips.

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Webinar
Considerations for High-Efficiency Genome Editing of Human Pluripotent Stem Cells
Webinar
Organoid Mini-Symposium: Utilization of Next Generation CRISPR Tools to Model Health and Disease in Adult Stem Cell-Derived Organoids
Webinar
Re-imagining CRISPR Technology for Use with Primary Immune Cells: New Insights for Cell-Based Immunotherapy Research
Webinar
Optimized Workflows for High-Efficiency Genome Editing in Stem and Primary Cell Types

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References

  1. Liang X et al. (2015) Rapid and highly efficient mammalian cell engineering via Cas9 protein transfection. J Biotechnol. 208: 44-53.

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